Project description:To investigate the role of a TIE1-function blocking antibody on gene expression of lung endothelial cells. Lung endothelial cells were FACS-isolated from primary tumor-bearing mice. Mice were treated either with IgG or TIE1-blocking antibody during primary tumor growth. Total RNA were used for microarray analysis using Clariom S mouse assays (Applied Biosystems).
Project description:Exaggerated signaling by vascular endothelial growth factor (VEGF) A and its receptor, VEGFR2, in pathologies results in poor vessel function. Still, pharmacological suppression of VEGFA/VEGFR2 may aggravate disease. Delineating VEGFR2 signaling in vivo provides strategies for suppression of specific VEGFR2-induced pathways. Three VEGFR2 tyrosine residues (Y949, Y1212, Y1173) induce downstream signaling. Here, we show that knock-in of phenylalanine to create VEGFR2 Y1212F in C57Bl/6 and FVB mouse strains leads to loss of growth factor receptor bound (GRB)2- and phosphoinositide 3´kinase (PI3K)p85-signaling. C57Bl/6 Vegfr2Y1212F/Y1212F show reduced embryonic endothelial cell (EC) proliferation and partial lethality. FVB Vegfr2Y1212F/Y1212F show reduced postnatal EC proliferation. Reduced EC proliferation in Vegfr2Y1212F/Y1212F explants is rescued by c-Myc overexpression. We conclude that VEGFR2 Y1212 signaling induces activation of extracellular-signal regulated kinase (ERK)1/2 and Akt pathways required for c-Myc-dependent gene regulation, endothelial proliferation and vessel stability. We use microarray to determine the differences in gene expression in endothelial cells between WT or Vegfr2Y1212F/Y1212F mice.
Project description:Following the identification of a critical time window of Blood Brain Barrier formation in the mouse embryo, we aimed to identify genes important for barriergenesis. To this end, we isolated cortical and lung E13.5 endothelial cells and compared expression between the two populations. The working hypothesis was that endothelial cells which are actively building a barrier would have a uniqe pattern of gene expression that would be detectable in comparison to a non-barrier endothelial population that is also active in vasculogenesis.
Project description:Cellular sources of liver endothelial cells remain elusive. Here, we used irradiation-conditioned bone marrow chimeric mice to lineage trace the endothelial cells. Bone marrow-derived (YFP+) and resident (YFP-) liver endothelial cells were isolated and total RNA were used for microarray analysis using Clariom S mouse assays (Applied Biosystems).
Project description:Following the identification of a critical time window of Blood Brain Barrier formation in the mouse embryo, we aimed to identify genes important for barriergenesis. To this end, we isolated cortical and lung E13.5 endothelial cells and compared expression between the two populations. The working hypothesis was that endothelial cells which are actively building a barrier would have a uniqe pattern of gene expression that would be detectable in comparison to a non-barrier endothelial population that is also active in vasculogenesis. E13.5 Tie2-GFP embryos were micro-dissected for cortex and lungs. Cortex tissue was carefully cleared of the meninges and choroid plexus. FACS purification of GFP positive cells and GeneChip analysis was applied . All material from a single litter (10-13 embryos) was pooled and considered as a biological replicate. n=4 litters.
Project description:Our goal is to study the contribution of endothelial, smooth muscle, and macrophage gene expression to early atherosclerosis using vessels from healthy, pre-lesioned, and lesioned mice. To accomplish this, we developed a method to isolate endothelial cells directly from the aorta and quantitate transcripts in RNA isolated from these cells. We performed expression array analysis on RNA isolated directly from the vascular cells of individual control, pre-lesioned, and atherosclerotic mouse aortas. Array expression analysis was not performed on smooth muscle cells or macrophages.